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Mol. Cell. Biol., Jan 1995, 315-325, Vol 15, No. 1
Copyright © 1995, American Society for Microbiology

A new platelet-derived growth factor-regulated genomic element which binds a serine/threonine phosphoprotein mediates induction of the slow immediate-early gene MCP-1

RR Freter, JA Alberta, KK Lam and CD Stiles
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts.

The MCP-1 chemokine gene belongs to a cohort of immediate-early genes that are induced with slower kinetics than c-fos. In this study, we identified a cluster of four platelet-derived growth factor (PDGF)- responsive elements within a 240-bp enhancer found in the distal 5' flanking MCP-1 sequences. Two of the elements bind one or more forms of the transcription factor NF-kappa B. We focused on the other two elements which are hitherto unreported, PDGF-regulated genomic motifs. One of these novel elements, detected as a 28-mer by DNase I footprinting, restores PDGF inducibility when added in two copies to a 5' truncated MCP-1 gene. A single copy of the second novel element, a 27-mer, restores PDGF inducibility to a 5' truncated MCP-1 gene. The 27- base element interacts with a PDGF-activated serine/threonine phosphoprotein that is detected only within the nucleus of PDGF-treated 3T3 cells. DNA binding of this phosphoprotein is activated by PDGF treatment with slow kinetics that match the time course of MCP-1 gene expression, and activation is not inhibited by cycloheximide. PDGF- activated binding to the 27-mer is shown to involve a single 30-kDa protein by UV-cross-linking analysis.


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